Routine Identification Research Articles

Hyperspectral Raman Imaging for Automated Recognition of Human Renal Amyloid.

In the clinical setting, routine identification of the main types of tissue amyloid deposits, light-chain amyloid (AL) and serum amyloid A (AA), is based on histochemical staining; rarer types of amyloid require mass spectrometry analysis. Raman spectroscopic imaging is an analytical tool, which can be used to chemically map, and thus characterize, the molecular composition of fluid and solid tissue. In this proof-of-concept study, we tested the feasibility of applying Raman spectroscopy combined with artificial intelligence to detect and characterize amyloid deposits in unstained frozen tissue sections from kidney biopsies with pathologic diagnosis of AL and AA amyloidosis and control biopsies with no amyloidosis (NA). Raman hyperspectral images, mapped in a 2D grid-like fashion over the tissue sections, were obtained. Three machine learning-assisted analysis models of the hyperspectral images could accurately distinguish AL (types λ and κ), AA, and NA 93-100% of the time. Although very preliminary, these findings illustrate the potential of Raman spectroscopy as a technique to identify, and possibly, subtype renal amyloidosis.

First Argentine database for the accurate identification of Brucella to species level by MALDI-TOF MS

MALDI-TOF mass spectrometry (MS) has proven to be a fast and reliable method for the identification of a large number of taxonomic groups. It offers the advantage of being able to incorporate protein spectra of microorganisms that are absent or poorly represented in commercial databases, such as the genus Brucella. The aim of the study was to build the first database of protein spectra of local biological variants of Brucella in Argentina and of standard strains. First, the identification performance of a panel of 135 strains was evaluated with the Swedish database ¨Folkhälsomyndigheten¨ (containing protein spectra of several international standards of the genus Brucella) imported from the open access site https://spectra.folkhalsomyndigheten.se/spectra/. With this library 100 % of the strains were correctly identified by mass spectrometry to genus level, but not to species level.Due to the limitation found, an in-house database was designed with local Brucella isolates from Argentina and standard strains used in routine bacteriological diagnosis. For its validation, a panel of strains, different from those used to develop the extended local database (n: 177), was used to, simultaneously, challenge both libraries. The samples were processed by triplicate and the results obtained were: 177 strains correctly identified to genus and species level compared to the gold standard method (phenotypic typing), meeting the criteria accepted by the literature and the manufacturer as reliable identification. Only 2 of these isolates had score values lower than 2 (1.862) and were therefore not included in the calculation of results.According to these results, MALDI-TOF MS is a fast and reliable method for the routine identification of the different Brucella species, and even has the advantage of reducing the time of exposure to pathogenic microorganisms for laboratorians. It could be considered a valuable technique to replace, in the near future, the current conventional techniques due to the ease of transferring protein spectra, avoiding the use of reference strains that are difficult to find commercially available and commonly used in phenotypic typing.

A-279 Comparison of Direct Identification and Short Incubation Time Methods of MALDI-TOF MS from Positive Blood Vials

Abstract Background MALDI-TOF analysis after a short incubation time of positive blood culture is currently one of the most widely used methods for rapidly identifying microorganisms. This method can identify most pathogenic bacteria within 4 to 6 h after blood culture. This experiment aims to use direct identification for MALDI-TOF analysis, which can shorten the identification time of bacteria and analyze the advantages and disadvantages of the two methods. Method We collected from January 2022 to December 2022, a total of 295 bottles of a single infection. After pretreatment methods such as removing cells and high-speed centrifugation to collect bacteria, carry out direct identification by MALDI-TOF then culture for 5 h for short incubation time method. The culture medium was continued incubated for 18–20 h, to confirm the final identification results of the strains and compare the identification accuracy of direct identification and short incubation time methods. Definition: confidence score cut-off values > = 1.7 of direct identification method. Result The identification rate of score > = 1.7 were 67%(199/295). Gram-negative bacilli were identified in 89%, followed by anaerobes in 60% and Gram-positive species in 56%. The yeast could not be identified either by the short incubation time method or the direct identification method. The short incubation time method identified 90% of Gram-negative bacilli, followed by 79% of Gram-positive bacteria and 7% of anaerobic bacteria. The rapid identification of anaerobic bacteria was significantly better than the identification results of the short incubation time method (60%:7%) (P value = 0.01). In addition, the results can be obtained within one hour by direct identification, which can save at least 5-fold times required for the short incubation time method of about 5 h, although the effect is slightly worse than short incubation time method (67 %:76%). Especially for Gram-negative bacilli, there is no statistically significant difference between direct identification and short incubation time methods; it proves that direct identification method can indeed replace short incubation time method. Conclusion Patients with septic shock who were treated with antibiotics within one hour had an 80% chance of survival; however, for every hour of delay within six hours of onset, the survival rate decreased by 7.6%. Compared with the short incubation time method, the direct identification method can report the correct strain to clinician 5-fold times earlier. This rapid, simplified extraction direct identification method is cost-effective, time-saving, and easy to use. The SSC sepsis guidelines in 2016 pointed out that extending the course of antibiotic treatment not only has no additional benefits but also is prone to drug resistance, resulting in side effects of antibiotics. Therefore, the direct identification method can significantly improve the optimal utilization of antibiotics for effective targeted treatment and reduce mortality, especially for the identification of anaerobic bacteria, which can provide faster identification results. Therefore, it is recommended that clinical laboratories include a direct identification method for routine identification of blood culture, and if they cannot be identified, they can be supplemented by a short incubation time method, to shorten the time of blood culture identification and effectively improve the treatment of bloodstream infections.

90 Screening Test Accuracy and Reliability of the Parent-Report Preschool Strengths and Difficulties Questionnaire (P-SDQ) in Primary Care Settings

Abstract Introduction/Background Previous research shows that mental health disorders can be identified in preschoolers. Detecting disorders early in life is key for early intervention and prevention of life-long behavioural and emotional issues. Individuals who experience mental health issues before the age of 14 years are at increased risk of a mental health disorder in adulthood, contrary to the idea that issues seen in young children are transient. Although the rate of mental health disorders in preschoolers is similar to the rate in older children, there is no agreed upon standard of care for routine screening or case identification of psychiatric problems in young children. Improving mental health and long-term outcomes for children requires early identification and monitoring of children at risk, including a screening tool that can be used in primary care settings. Objectives To evaluate the criterion validity (screening test accuracy) and test-retest reliability of the parent-report preschool Strengths and Difficulties Questionnaire (P-SDQ) in primary care settings for a sample of 2-4 year olds. Specifically, we were interested in evaluating the screening test accuracy of the P-SDQ for presence of DSM-5 diagnoses. Design/Methods Children 24-48 months were recruited at scheduled primary care visits in Toronto, Canada. Parents completed the P-SDQ at baseline, 2 and 12 weeks. At 12 weeks, parents participated in the Preschool Age Psychiatric Assessment (PAPA). We assessed the criterion validity of the baseline P-SDQ using three different criterion measures. We compared the Total Difficulties Score (TDS) with presence of any DSM-5 diagnoses on the PAPA. For the internalizing subscale score, the criterion was presence of any internalizing disorder. For the externalizing subscale score, the criterion was presence of any externalizing disorder. Criterion validity was evaluated using area under the curve (AUC) and calculating screening test properties for a range of threshold values of the TDS and two subscale scores. Optimal thresholds were selected based on the maximal value of both sensitivity and specificity. Test-retest reliability at baseline and 2 weeks was evaluated using intraclass correlation coefficient (ICC). Results 183 children were enrolled, mean age 39.3 (SD 7.4) months, 46.4% male, 120 (66%) completed the P-SDQ at 2 weeks, 107 (58%) completed the PAPA at 12 weeks. Of those with a PAPA, 26 (24%) had any psychiatric diagnosis, 22 (21%) had an internalizing disorder, 9 (8%) had an externalizing disorder, and 5 (5%) had both. TDS identified any diagnosis with AUC = 0.67 (95% CI: 0.55, 0.79); the internalizing subscale identified internalizing disorders with AUC = 0.61 (95% CI: 0.47, 0.74); the externalizing subscale identified externalizing disorders with AUC = 0.77 (95% CI: 0.60, 0.94). Sensitivity and specificity for TDS were 50% and 78%; for internalizing subscale were 18% and 99%; for externalizing subscale were 78% and 71%. Test-retest reliability (ICC) for the TDS was 0.72 (95% CI: 0.62, 0.80); internalizing subscale was 0.62 (95% CI: 0.49, 0.71); externalizing subscale was 0.79 (95% CI: 0.72, 0.86). Conclusion The parent report P-SDQ externalizing subscale has sufficient accuracy and reliability for identifying externalizing disorders in primary care. The TDS and internalizing subscale perform less well. Table 3: Screening test properties of the P-SDQ compared with the PAPA (n=107)

Processes for identifying caregivers and screening for caregiver and patient distress in community oncology: results from WF-1803CD.

Despite their vital roles, informal caregivers of adult cancer patients are commonly overlooked in cancer care. This study describes processes for identifying cancer caregivers and processes for distress screening and management among caregivers and patients in the understudied community oncology setting. Supportive care leaders from the National Cancer Institute Community Oncology Research Program practices completed online survey questions regarding caregiver identification, caregiver and patient distress screening, and distress management strategies. We described practice group characteristics and prevalence of study outcomes. Multivariable logistic regression explored associations between practice group characteristics and caregiver identification in the electronic health record (EHR). Most (64.9%, 72 of 111) supportive care leaders reported routine identification and documentation of informal caregivers; 63.8% record this information in the EHR. Only 16% routinely screen caregivers for distress, though 92.5% screen patients. Distress management strategies for caregivers and patients are widely available, yet only 12.6% are routinely identified and screened and had at least 1 referral strategy for caregivers with distress; 90.6% are routinely screened and had at least 1 referral strategy for patients. Practices with a free-standing outpatient clinic (odds ratio [OR] = 0.29, P = .0106) and academic affiliation (OR = 0.01, P = .04) were less likely to identify and document caregivers in the EHR. However, higher oncologist volume was associated with an increased likelihood of recording caregiver information in the EHR (OR = 1.04, P = .02). Despite high levels of patient distress screening and management, few practices provide comprehensive caregiver engagement practices. Existing patient engagement protocols may provide a promising platform to build capacity to better address caregiver needs.

Unexpected mapping of recurrent laryngeal nerve by fluorescence-guided surgery using near-infrared indocyanine green angiography

Background. The recurrent laryngeal nerve (RLN) damage and parathyroid gland injury are the most severe complications of thyroid surgery. The possibility of RLN confirmation in the near-infrared spectrum after the injection of indocyanine green (ICG) was not yet been studied. Aim: to evaluate the ICG angiography for the identification of RLN during thyroid and parathyroid surgery. Materials and methods. ICG angiography of RLN was performed in 7 patients. An intraoperative neuromonitoring was applied as a method of controlling RLN. During the operation, parathyroid glands and RLN were identified by visual inspection (naked eye). To further confirm the location of the parathyroid glands by autofluorescence, an intravenous injection of ICG was performed with a concentration of 0.25 mg/kg followed by the application of the image-based system. Results. A good signal was achieved in the near-infrared spectrum from the RLN in all cases after the ICG injection. Sufficient blood perfusion of the RLN could be considered as a reasonable explanation for the exhibition of a good ICG near-infrared signal. Conclusions. ICG use might be a helpful approach for the confirmation of the RLN in addition to routine visual identification. Such function could be applied during fluorescence-guided surgery to evaluate the parathyroid gland autofluorescence. Visualization of RLN by ICG angiography is considered as an additional useful feature to prevent RLN injury.

Routine identification of children of adults with mental illness: a nationwide survey of healthcare practitioners’ perspectives and practices in Japan

ABSTRACT Objective The association was investigated between healthcare practitioners’ actions of routinely identifying children of adults with mental illness by asking whether they have children under the age of 18 and practitioners’ perspectives and practices regarding supporting parents and their children. The factors that contribute to practitioners’ such routine identification were also determined. Method A cross-sectional nationwide survey was conducted using an anonymous self-report questionnaire among Japanese healthcare practitioners. Results The responses (84.7% were nurses) from 1138 participants were analysed. More than half of the participants (73.7%) routinely identified whether adults with mental illness had children. Participants who routinely identified children also provided more support to children and to the client regarding his or her parenting. Three factors were found to contribute to the participants’ routine identification of children: (1) recognising ‘difficulty with parenting' as a reason why support for children is necessary; (2) ‘assessing parenting status when supporting a parent with a mental illness' and (3) realising that participants ‘do not know how to provide support' as a reason why support for children is difficult. Discussion Providing education to practitioners regarding supporting children whose parents have a mental illness will help increase the routine identification of children in mental health services.

Spectrometric Characterization of Clinical and Environmental Isolates of Aspergillus Series Versicolores.

Aspergillus series Versicolores are molds distributed among 17 species, commonly found in our environment, and responsible for infections. Since 2022, a new taxonomy has grouped them into 4 major lineages: A. versicolor, A. subversicolor, A. sydowii, and A. creber. Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) could be a faster and more cost-effective alternative to molecular techniques for identifying them by developing a local database. To evaluate this technique, 30 isolates from Aspergillus series Versicolores were used. A total of 59 main spectra profiles (MSPs) were created in the local database. This protocol enabled accurate identification of 100% of the extracted isolates, of which 97% (29/30) were correctly identified with a log score ≥ 2.00. Some MSPs recorded as Aspergillus versicolor in the supplier's database could lead to false identifications as they did not match with the correct lineages. Although the local database is still limited in the number and diversity of species of Aspergillus series Versicolores, it is sufficiently effective for correct lineage identification according to the latest taxonomic revision, and better than the MALDI-TOF MS supplier's database. This technology could improve the speed and accuracy of routine fungal identification for these species.

Cleft Lip and Cleft Palate: Time to Include Orofacial Ultrasound Markers Into the First-Trimester Anatomy Scan?

Orofacial clefts are one of the most common congenital malformations. The prenatal diagnosis is often made in the second trimester of pregnancy as result of ultrasound examination of the midface on coronal and axial planes. However, the diagnosis in the first trimester is elusive due to the small size of the facial structures and technical limitations present at this early gestational age. In this Commentary, we suggest the routine systematic ultrasound identification of easy-to-obtain landmarks to improve the detection of cleft lip and cleft palate in the first trimester. These include, but are not limited to, visualization of the primary palate using the coronal plane of the face looking for disruption at the base of the retronasal triangle, and visualization of the palate using the sagittal plane looking for the maxillary gap and loss of the superimposed line. Early prenatal detection of orofacial clefts would allow a more detailed search for associated chromosomal anomalies or genetic syndromes.

Evaluation of the Zybio EXS3000 mass spectrometry in routine identification of Clinical isolates

The matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been widely applied in routine clinical microbiology laboratories as an efficient and reliable technique for diagnostic purpose. In this work, we evaluated the performance of the newly developed Zybio EXS3000 (Zybio Inc., China) in microbial identification and compared it with VITEK MS (bioMérieux, France). For this study, a total of 1340 isolates from various clinical specimens were collected. These isolates were analyzed simultaneously on both EXS3000 and VITEK MS. The inconsistent or unidentifiable data were further identified using the help of either 16S rRNA gene or ITS region sequencing. During the study, we observed that EXS3000 and VITEK MS provided positive confirmatory diagnostics for 95.0% and 96.5% of the isolates, respectively, which were consistent with the sequencing results. However, it is worth noting that the EXS3000 system needs to improve the identification performance of Candida albicans in the follow-up. There are no significant differences between the two devices in terms of microbial identification performance. The advantage of EXS3000 over VITEK MS is in its ability to perform in significantly lesser time period. In conclusion, the results of this investigation showed that EXS3000 can be used to identify microorganisms in clinical microbiology laboratories.

Machine learning analysis of wing venation patterns accurately identifies Sarcophagidae, Calliphoridae and Muscidae fly species.

In medical, veterinary and forensic entomology, the ease and affordability of image data acquisition have resulted in whole-image analysis becoming an invaluable approach for species identification. Krawtchouk moment invariants are a classical mathematical transformation that can extract local features from an image, thus allowing subtle species-specific biological variations to be accentuated for subsequent analyses. We extracted Krawtchouk moment invariant features from binarised wing images of 759 male fly specimens from the Calliphoridae, Sarcophagidae and Muscidae families (13 species and a species variant). Subsequently, we trained the Generalized, Unbiased, Interaction Detection and Estimation random forests classifier using linear discriminants derived from these features and inferred the species identity of specimens from the test samples. Fivefold cross-validation results show a 98.56 ± 0.38% (standard error) mean identification accuracy at the family level and a 91.04 ± 1.33% mean identification accuracy at the species level. The mean F1-score of 0.89 ± 0.02 reflects good balance of precision and recall properties of the model. The present study consolidates findings from previous small pilot studies of the usefulness of wing venation patterns for inferring species identities. Thus, the stage is set for the development of a mature data analytic ecosystem for routine computer image-based identification of fly species that are of medical, veterinary and forensic importance.

Making Trauma Less Traumatic: Implementing Inpatient Pediatric Screening of Acute Stress Symptoms.

Pediatric patients hospitalized for physical injury and trauma are at increased risk of developing acute stress disorder. Routine identification and referral for treatment of patients with psychological sequelae of traumatic injury were added to the accreditation requirements of the American College of Surgeons for pediatric trauma centers in 2022. We aimed to use quality improvement methodology with iterative interventions to increase psychological screening consults for admitted pediatric trauma patients to 80% in 6 months. We planned a quality improvement intervention to increase identification of posttraumatic psychological symptoms in pediatric trauma patients. We created a Making Trauma Less Traumatic consultation service with a dedicated therapist to provide screening, treatment, and referral. Key interventions included education of key stakeholders and iterative improvements in consultation workflow. Our primary measure was frequency of eligible pediatric trauma patients who had consultation requests for posttrauma therapy during admission. We additionally monitored percent of patients with positive symptom screens and lost to follow-up. From September 2020 through November 2021, consults for eligible pediatric trauma patients improved from a baseline of 4.1% to a weekly mean of 100%. Of those screened, 32.7% had at least 1 symptom of acute stress. No families declined screening or therapy, though 29.5% were lost to intended follow-up. We present a successful model of implementing routine psychological screening of pediatric trauma patients utilizing a dedicated consultation service. A high number of admitted patients screened positive for symptoms during hospitalization. Families were accepting of the intervention though follow-up was challenging.

Highly informative SNP markers for routine individual identification and certification of forest reproductive material in Quercus ilex and Q. suber and their hybrids

Holm oak (Quercus ilex L.) and cork oak (Q. suber L.) are two species of great ecological and socio-economical relevance in the Mediterranean western basin, mainly in the Iberian Peninsula. However, although breeding programs are in progress, basic materials providing qualified and tested Forest Reproductive Materials (FRM) are still lacking. In order to enable an accurate identification at the individual level, allowing the certification of selected material, efficient and affordable molecular markers are required.We report the identification of different sets of 96 and 500 single nucleotide polymorphism markers, selected in order to: I) allow the unambiguous identification at the individual level, both for holm oak and for cork oak and II) provide an accurate estimation of introgression level in hybrid individuals, at a low genotyping cost. These molecular tools fulfill the requirements for FRM certification, as well as for research and breeding purposes.

Correlation between visual scores and parathyroid function.

This mini review summarizes the controversies regarding routine parathyroid identification reviews publications that assess visual scores to predict parathyroid function after thyroidectomy during the pre-ICG era.

Розлади ендотеліальної функції в дітей, які перехворіли на інфекцію COVID-19

Purpose - to learn the features of clinical condition and state of endothelial function as the marker of the development of cardiovascular pathology in children who suffered from COVID-19. Materials and methods. The research group consisted of 70 children aged 7-14 without chronic pathology who suffered from COVID-19 and had laboratory confirmation of the disease. Assessment of the functioning of the vessels’ endothelium was done by studying the dynamics of blood flow in the brachial artery and changes of it diameter at rest and during reactive hyperemia after an occlusion test. Statistical assessment of the obtained data was carried out by the application package «Statistica 10.0 for Windows» using the method of variational statistics. Student’s t-test was calculated to assess the reliability of mean values differences. Results. The state of endothelial function show that 85.4% of children after COVID-19 infection had signs of endothelial dysfunction, mainly hypoergic (32.9%) and paradoxical (30%) forms, while normoergic function of the endothelium was found only in 14.6% of children. In the same time in the group of children who did not suffer from COVID-19, was found that the normorergic form registrated in 80.0%, hyperergic endothelial dysfunction was found in 13.3% of children, and hypoergic - in 6.7% of children. Paradoxical endothelial dysfunction was not found in any child from this group. Conclusions. COVID-19 infection has negative consequences on the endothelium function and contributes to the development of endothelial dysfunction regardless of the severity of the disease course. Our results obtained the need of future study to develop an affordable and low-cost algorithm for routine use and identification of children with markers of lesion of cardiovascular system. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

MALDI-TOF MS-Based KPC Direct Detection from Patients' Positive Blood Culture Bottles, Short-Term Cultures, and Colonies at the Hospital.

Carbapenemase resistance in Enterobacterales is a global public health problem and rapid and effective methods for detecting these resistance mechanisms are needed urgently. Our aim was to evaluate the performance of a MALDI-TOF MS-based "Klebsiella pneumoniae carbapenemase" (KPC) detection protocol from patients' positive blood cultures, short-term cultures, and colonies in healthcare settings. Bacterial identification and KPC detection were achieved after protein extraction with organic solvents and target spot loading with suitable organic matrices. The confirmation of KPC production was performed using susceptibility tests and blaKPC amplification using PCR and sequencing. The KPC direct detection (KPC peak at approximately 28.681 Da) from patients' positive blood cultures, short-term cultures, and colonies, once bacterial identification was achieved, showed an overall sensibility and specificity of 100% (CI95: [95%, 100%] and CI95: [99%, 100%], respectively). The concordance between hospital routine bacterial identification protocol and identification using this new methodology from the same extract used for KPC detection was ≥92%. This study represents the pioneering effort to directly detect KPC using MALDI-TOF MS technology, conducted on patient-derived samples obtained from hospitals for validation purposes, in a multi-resistance global context that requires concrete actions to preserve the available therapeutic options and reduce the spread of antibiotic resistance markers.

#4186 FEASIBILITY OF NEPHROLOGY-TAILORED GERIATRIC ASSESSMENT: AN IMPLEMENTATION STUDY

Abstract Background and Aims Geriatric evaluation of older patients with progressive chronic kidney disease is recommended in guidelines to enhance decision-making for kidney replacement choices. Although frailty, physical and cognitive impairments, and depression are prevalent, routine identification of geriatric impairments is lacking. Therefore, we aimed to study feasibility and determinants of successful implementation of a novel approach: nephrology-tailored geriatric assessment. Method In a mixed-methods implementation study, we investigated the feasibility of implementation of a consensus-based geriatric assessment [1] in Dutch nephrology centers. Feasibility was assessed through hospital participation and patient inclusion (among patients aged 70+ years with eGFR≤20mL/min/1.73 m2). We also evaluated determinants of successful implementation with a post-implementation survey among healthcare professionals covering (i) perceived enablers and barriers of implementation, (ii) perceived integration in work routines (Normalization MeAsure Development tool), and (iii) perceived relevance of the instruments. In addition, patients were asked if the geriatric assessment provided them with new insights for their future treatment decision for kidney failure. Results Feasibility was substantiated in implementation of geriatric assessment in 10 hospitals among 191 patients, i.e. 100% and 96% of our aimed goals. According to participating healthcare professionals (n = 71), determinants that facilitated implementation were: collaboration with the geriatric department, multidisciplinary meetings and reports, and execution of assessments by nurses. Most frequently mentioned barriers to implementation were: patient illiteracy or language barrier, lack of time, and burden for patients. Professionals considered geriatric assessment sufficiently integrated into work routines (mean 6.7 of 10, SD 2.0), but also subject to improvement (Figure 1). Implementation of geriatric assessment was considered sufficiently relevant for routine care (mean score 7.8 of 10, SD 1.2). Half of the patients (n = 42/82) stated that the geriatric assessment provided them with new insights, whereas 27% (n = 22) responded not to have gained new insights from the tests. Conclusion Multicenter implementation of nephrology-tailored geriatric assessment seems feasible and relevant. For future implementation we recommend involving the geriatric department, using multidisciplinary teams and -meetings, acknowledging the pivotal role of nurse practitioners, and using a flexible approach matching local resources.

Pathogenic bacteria recovered from Gene X-pert tuberculosis-negative adult patients in Gondar, Northwest Ethiopia

IntroductionLower respiratory tract infections (LRTIs) caused by drug-resistant pathogenic bacteria is a major problem in developing countries including Ethiopia. Therefore, this study aimed to determine the pathogenic bacteria and their antimicrobial susceptibility patterns among Gene X-pert tuberculosis-negative adult patients with clinically suspected LRTIs at the University of Gondar Comprehensive Specialized Referral Hospital, Gondar, Northwest Ethiopia.MethodsThis institutional-based cross-sectional study was conducted from February 01 to March 15, 2020. Socio-demographic data were collected by using a structured questionnaire. A total of 254 sputum specimens were collected from Gene X-pert tuberculosis-negative patients. Bacterial recovery was performed using blood, chocolate, and MacConkey agar plates. Bacterial isolates were identified based on Gram staining, colony characteristics, and biochemical reactions. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method. Methicillin resistance of S. aureus was confirmed using cefoxitin (30 µg). Descriptive statistics were calculated for each variable and results are shown in tables and figures.ResultsIn this study, the overall sputum culture positivity rate was 145/254 (57.1%). Gram-negative bacteria 111 (64.9%) were predominant compared to Gram-positive bacteria 60 (35.1%). Of the 145 culture-positive cases, 26 (14.8%) had poly-bacterial infections. S. aureus 40 (66.7%) was the predominant Gram-positive bacterium whereas K. pneumoniae 33 (29.7%), was the most isolated Gram-negative bacterium. Bacterial species, such as S. aureus were sensitive to ciprofloxacin 38/40 (95.0%), gentamicin 37/40 (92.5%), cefoxitin 36/40 (90.0%), and clindamycin 34/40 (85.0%). The proportion of Methicillin-resistant S. aureus was low, 4(10.0%). S. pneumoniae was sensitive to chloramphenicol 8/9 (88.9%) and resistant to ciprofloxacin 6/9 (66.7%). K pneumoniae, P. aeruginosa, E. coli, Serratia species, and H. influenzae also demonstrated high levels of resistance to ampicillin at rates of 21/33 (63.6%), 8/8 (100.0%), 15/17 (88.2%), 7/10 (70.0%), and 6/6 (100.0%), respectively.ConclusionThis study revealed a higher burden of Gram-negative and Gram-positive pathogenic bacterial agents, which is responsible for LRTs. Therefore, routine sputum culture identification and antibiotic susceptibility testing should be performed in Gene X-pert tuberculosis-negative patients.

Geographical origin identification of Chinese red wines using ultraviolet-visible spectroscopy coupled with machine learning techniques

Identifying geographical origins of red wines produced in specific regions is of great importance, since the geographical origins of wine influence its quality and price greatly. In this study, the feasibility of UV-Vis spectroscopy was evaluated for the classification of Chinese red wine samples according to their geographical origins, using principal component analysis (PCA) and two machine learning techniques: orthogonal partial least squares-discriminant analysis (OPLS-DA) and support vector machine (SVM). The PCA analysis indicated that there are differences in the chemical composition between wine samples from three different origins and inferred the chemical compounds responsible for the discrimination between wine geographical origins. Furthermore, OPLS-DA and SVM models were established to predict the class membership of wine samples from three different origins and results showed that both models can provide correct recognition rates of 100 % for wine samples in training and prediction sets. This study demonstrated that the combination of UV-Vis spectroscopy with machine learning-based modeling has the potential to be a simple, fast and low-cost tool for the routine identification of geographical origins of Chinese red wines.

Differential Detection of Alternaria alternata Haplotypes Isolated from Carya illinoinensis Using PCR-RFLP Analysis of Alt a1 Gene Region.

Alternaria black spot disease on pecan is caused by the opportunistic pathogen Alternaria alternata and poses a serious threat to the local South African and global pecan industry. Several diagnostic molecular marker applications have been established and used in the screening of various fungal diseases worldwide. The present study investigated the potential for polymorphism within samples of A. alternata isolates obtained from eight different geographical locations in South Africa. Pecan (Carya illinoinensis) leaves, shoots, and nuts-in-shuck with Alternaria black spot disease were sampled, and 222 A. alternata isolates were retrieved. For rapid screening to identify Alternaria black spot pathogens, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the Alternaria major allergen (Alt a1) gene region was used, followed by the digestion of the amplicons with HaeIII and HinfI endonucleases. The assay resulted in five (HaeIII) and two (HinfI) band patterns. Unique banding patterns from the two endonucleases showed the best profile and isolates were grouped into six clusters using a UPGMA (unweighted pair group method with arithmetic averages) distance matrix (Euclidean) dendrogram method on R-Studio. The analysis confirmed that the genetic diversity of A. alternata does not depend on host tissues or the pecan cultivation region. The grouping of selected isolates was confirmed by DNA sequence analysis. The Alt a1 phylogeny corroborated no speciation within the dendrogram groups and showed 98-100% bootstrap similarity. This study reports the first documented rapid and reliable technique for routine screening identification of pathogens causing Alternaria black spot in South Africa.